Corn plant and seed corresponding to transgenic event MON89034 and methods for detection and use thereof

ABSTRACT

The present invention provides a transgenic corn event MON89034, and cells, seeds, and plants comprising DNA diagnostic for the corn event. The invention also provides compositions comprising nucleotide sequences that are diagnostic for said corn event in a sample, methods for detecting the presence of said corn event nucleotide sequences in a sample, probes and primers for use in detecting nucleotide sequences that are diagnostic for the presence of said corn event in a sample, growing the seeds of such corn event into corn plants, and breeding to produce corn plants comprising DNA diagnostic for the corn event.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.14/078,275, filed Nov. 12, 2013, which application is a divisional ofU.S. patent application Ser. No. 13/114,022, filed May 23, 2011, nowissued as U.S. Pat. No. 8,581,047, which application is a continuationof US patent application Ser. No. 11/753,574, filed May 25, 2007, nowissued as U.S. Pat. No. 8,062,840, which claims the benefit of priorityto U.S. Provisional Application Ser. No. 60/808,834, filed May 26, 2006,each of the disclosures of which are incorporated herein by reference intheir entirety.

FIELD OF THE INVENTION

The present invention relates to transgenic corn event MON89034 andplant parts and seed thereof. The event exhibits resistance to insectinfestation from insects in the order Lepidoptera. The invention alsorelates to methods for using plants and seeds comprising DNA that isdiagnostic for the presence of the transgenic event when probed for thepresence of nucleotide sequences that are unique to the transgenicevent, and to methods for detecting the presence of said corn event in abiological sample by detecting specific nucleotide sequences that areunique to the transgenic event. The invention provides nucleotidesequences that are unique to the event.

BACKGROUND OF THE INVENTION

This invention relates to the Lepidopteran resistant transgenic varietyof corn (Zea mays) plant referred to herein as event MON89034, and tounique DNA sequences present that, when detected in any sample orvariety of corn, are diagnostic for the presence of the transgenic cornplant event MON89034 in that sample or variety, and also relates to thedetection of the transgene/genomic insertion region in corn MON89034,and progeny plants and seeds derived therefrom.

The corn plant event MON89034 is particularly resistant to insects inthe Lepidoptera family such as Fall armyworm (Spodoptera frugiperda),European corn borer (Ostrinia nubilalis), corn earworm (Helicoverpazea), southwestern corn borer (Diatraea grandiosella), and black cutworm(Agrotis ipsilon) and the like, all of which are agronomically importantinsect pests.

Corn is an important crop and is a primary food source in many areas ofthe world. Biotechnology methods have been applied to corn for thepurpose of improving agronomic traits and the quality of the product.One such agronomic trait is insect resistance, for example, geneticallyengineered resistance to lepidopteran and coleopteran species thatarises in corn plants genetically engineered to contain one or moregenes encoding insecticidal agents (see for example, U.S. Pat. No.6,489,542 and U.S. Pat. No. 6,620,988). It is advantageous to detect thepresence of a particular transgenic event in a biological sample inorder to determine whether one or more progeny of a sexual crosscontains the transgenic material. For example, the detection of theevent in a sample is important for licensing purposes, for establishingand maintaining purity standards, important for complying withregulatory agencies, for complying with food ingredient standards, foruse in legal proceedings in establishing that one or more particularindividuals or entities has been using the particular event without alicense from the owner or licensee of any patents directed to thetransgenic event, and for insuring compliance with various governmentregulations and/or laws.

In addition, methods that enable the detection of a particular plantwould be helpful when complying with regulations requiring thepre-market approval and labeling of foods derived from the recombinantcrop plants. Individuals or entities that are resistant to the presenceof a transgenic event in a sample also desire reliable methods fordetecting the presence of the transgene in a sample in order for them tobe able to capitalize on their business, which takes advantage of anabsence of transgenes in their products.

Despite these advantages, it is possible that insects may evolveresistance to plants expressing only one B. thuringiensis δ-endotoxin.Such resistance, should it become widespread, would clearly limit thecommercial value of germplasm containing single Bt genes.

One possible way of increasing the effectiveness of insecticidal agentsprovided via transgenic plants and directed at controlling target insectpests and contemporaneously reducing the likelihood of emergence ofinsect pests resistant to such insecticidal agents would be to ensurethat transgenic crops express high levels of these insecticidal agents,such as Bacillus thuringiensis delta-endotoxins (McGaughey and Whalon(1992), Science 258:1451-55; Roush (1994) Biocontrol. Sci. Technol.4:501-516). In addition, having a repository of insecticidal genes thatare effective against groups of insect pests and which manifest theireffects through different modes of action can safeguard againstdevelopment of resistance. The onset of resistance could besubstantially delayed as a result of providing a crop that expresses twoor more insecticidal activities exhibiting overlapping toxicity to thesame insect species. One means for achieving such dual modes of actioncould be to provide a plant expressing a Bt gene toxic to a particularinsect species along with a dsRNA that is provided for the purpose oftargeting for suppression an essential gene of the same insect speciestargeted by the Bt toxin, the dsRNA eliciting an RNAi response uponingestion by the target pest, providing a means for redundancy in theevent that the insect develops resistance either to the dsRNA or to theBt gene. Alternatively, co-expression in a plant of two or moreinsecticidal toxins both toxic to the same insect species but eachexhibiting a different mode of effectuating its killing activity,particularly when both are expressed at high levels, provides a meansfor effective resistance management. Examples of such insecticidesuseful in such combinations include but are not limited to Bt toxins,Xenorhabdus sp. or Photorhabdus sp. insecticidal proteins,deallergenized and de-glycosylated patatin proteins and/or permuteins,plant lectins, and the like.

The expression of foreign genes in plants is known to be influenced bytheir chromosomal position, perhaps due to chromatin structure (e.g.,heterochromatin) or the proximity of transcriptional regulation elements(e.g., enhancers) close to the integration site (Weising et al. (19880Ann. Rev. Genet 22:421-477). For this reason, it is often necessary toscreen a large number of events in order to identify an eventcharacterized by optimal expression of an introduced gene of interest.Even then, with dozens or even hundreds of different transgenic eventsin hand, there is no certainty of success in identifying a singletransgenic event that provides the optimum levels of expression of theat least two different toxins or insecticidal agents and lacks anyundesirable agronomic deficiencies or phytotoxic effects, either as aresult of the insertion into some essential or partially essentialregion of the plant genome, or as a result of toxic effects broughtabout by the levels of expression of the transgenes. For example, it hasbeen observed in plants and in other organisms that there may be widevariation in the levels of expression of an introduced gene amongevents. There may also be differences in spatial or temporal patterns ofexpression, for example, differences in the relative expression of atransgene in various plant tissues, that may not correspond to thepatterns expected from transcriptional regulatory elements present inthe introduced gene construct. For this reason, it is common to produceseveral hundreds to several thousands different events and screen theevents for a single event that has the desired transgene expressionlevels and patterns for commercial purposes. An event that has thedesired levels or patterns of transgene expression is useful forintrogressing the transgene into other genetic backgrounds by sexualoutcrossing using conventional breeding methods. Progeny of such crossesmaintain the transgene expression characteristics of the originaltransformant. This strategy is used to ensure reliable gene expressionin a number of varieties that are suitably adapted to specific localgrowing conditions.

It is possible to detect the presence of a transgene by any well knownnucleic acid detection method such as the polymerase chain reaction(PCR) or DNA hybridization using nucleic acid probes. These detectionmethods generally focus on frequently used genetic elements, such aspromoters, terminators, marker genes, or even the coding sequenceencoding the protein or dsRNA of interest expressed from thetransgene(s), etc. As a result, such methods may not be useful fordiscriminating between different events, particularly those producedusing the same DNA construct, unless the sequence of chromosomal DNAadjacent to the inserted DNA (“flanking DNA”) is known. Depending on themethod used for introducing the transgene(s) into a plant genome,abberant or unusual effects can be observed, which often severelycomplicate the identification of the plant genome sequences flanking thetransgenic DNA that was intended to be introduced into the plant. Often,rearrangements of the inserted DNA, rearrangements of the flankinggenome DNA, or rearrangements of both the inserted DNA and the flankinggenome DNA are prevalent, and complicate the analysis of the insertionalevent being evaluated. Therefore, it is advantageous to have a means forselecting, for identifying, and for insuring the purity andcharacteristics of a particular transgenic event in a sample, and theonly way to accomplish this is to identify one or more unique sequencesassociated only with the desired transgenic event, and the presence ofsuch sequences in a biological sample containing DNA of the plantspecies into which the transgenic DNA was inserted to give rise to theevent are thus diagnostic for the event in such sample.

SUMMARY OF THE INVENTION

The present invention is related to the transgenic corn plant designatedMON89034 and progeny that are indistinguishable from corn event MON89034(to the extent that they also contain at least one allele thatcorresponds to the inserted transgenic DNA) thereof having seeddeposited on Mar. 28, 2006 with American Type Culture Collection (ATCC)with Accession No. PTA-7455. Another aspect of the invention is theprogeny plants, or seeds, or regenerable parts of the plants and seedsof the corn event MON89034 that contain a polynucleotide selected fromthe group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ IDNO:4, and SEQ ID NO:5. The invention also includes plant parts of thecorn event MON89034 that include, but are not limited to pollen, ovule,flowers, shoots, roots, stalks, silks, tassels, ears, and leaves, solong as these parts contain at least the polynucleotides as set forthabove. Novel genetic compositions contained in the genome of MON89034and products such from MON89034 such as meal, flour, oil, pulp, andbiomass left over in a field of corn plants corresponding to MON89034event are an aspect of this invention.

The invention provides an insect resistant corn plant that has all ofthe physiological and morphological characteristics of the corn eventMON89034.

According to one aspect of the invention, compositions and methods areprovided for detecting the presence of the transgene/genomic insertionregion from a novel corn plant designated MON89034. DNA sequences areprovided that comprise at least one junction sequence of MON89034selected from the group consisting of SEQ ID NO:1 (located at positions2051 to 2070 on SEQ ID NO: 5) and SEQ ID NO:2 (located at positions11367 to 11388) and complements thereof; wherein a junction sequencespans the junction between heterologous DNA inserted into the genome andthe DNA from the corn cell flanking the insertion site and is diagnosticfor the event (FIG. 1). A corn event MON89034 and seed comprising theseDNA molecules is an aspect of this invention.

DNA sequences that comprise the novel transgene/genomic insertionregion, SEQ ID NO:3 and SEQ ID NO:4 (FIG. 2) from corn event MON89034are aspects of this invention. The corn plant and seed comprising thesemolecules are also aspects of this invention.

According to another aspect of the invention, two DNA molecules areprovided for use in a DNA detection method, wherein the first DNAmolecule comprises at least 11 or more contiguous polynucleotides of anyportion of the transgene region of the DNA molecule of SEQ ID NO:3 and aDNA molecule of similar length of any portion of a 5′ flanking corngenomic DNA region of SEQ ID NO:3, where these DNA molecules when usedtogether are useful as DNA primers in a DNA amplification method thatproduces an amplicon. The amplicon produced using these DNA primers inthe DNA amplification method is diagnostic for corn event MON 89304 whenthe amplicon contains SEQ ID NO:1. Any amplicon produced by DNA primershomologous or complementary to any portion of SEQ ID NO:3 and anyamplicon that comprises SEQ ID NO:1 is an aspect of the invention.

According to another aspect of the invention, two DNA molecules areprovided for use in a DNA detection method, wherein the first DNAmolecule comprises at least 11 or more contiguous polynucleotides of anyportion of the transgene region of the DNA molecule of SEQ ID NO:4 and aDNA molecule of similar length of any portion of a 3′ flanking corngenomic DNA of SEQ ID NO:4, where these DNA molecules are useful as DNAprimers in a DNA amplification method. The amplicon produced using theseDNA primers in the DNA amplification method is diagnostic for corn eventMON 89304 when the amplicon contains SEQ ID NO:2. Any amplicons producedby DNA primers homologous or complementary to any portion of SEQ ID NO:4and any amplicon that comprises SEQ ID NO:2 is an aspect of theinvention.

According to another aspect of the invention, methods of detecting thepresence of DNA corresponding to the corn event MON89034 in a sample areprovided. Such methods comprise: (a) contacting the sample comprisingDNA with a primer set that, when used in a nucleic acid amplificationreaction with genomic DNA from corn event MON89034, produces an ampliconthat is diagnostic for corn event MON89034; (b) performing a nucleicacid amplification reaction, thereby producing the amplicon; and (c)detecting the amplicon wherein said amplicon comprises SEQ ID NO:1 orSEQ ID NO:2.

A corn plant, or seed, or product derived from the plant or seedMON89034 wherein the genomic DNA comprising comprises a DNA moleculeconsisting essentially of SEQ ID NO:5 and complements thereof. A cornplant, or seed, or product derived from the plant or seed MON89034, inwhich the genomic DNA when isolated from the corn plant, or seed, orproduct comprises a DNA molecule incorporating nucleotides 2061 to 11377of SEQ ID NO:5 and complements thereof.

A corn plant, or seed, or product derived from the plant or seedMON89034, in which the genomic DNA when isolated from the corn plant, orseed, or product produces an amplicon in a DNA amplification method,wherein DNA primer molecules SEQ ID NO:6 and SEQ ID NO:7 is used in theDNA amplification method.

According to another aspect of the invention, methods of detecting thepresence of a DNA corresponding to the MON89034 event in a sample, suchmethods comprising: (a) contacting the sample comprising DNA with aprobe that hybridizes under stringent hybridization conditions withgenomic DNA from corn event MON89034 and does not hybridize under thestringent hybridization conditions with a control corn plant; (b)subjecting the sample and probe to stringent hybridization conditions;and (c) detecting hybridization of the probe to the corn event MON89034DNA wherein said probe comprises SEQ ID NO:1 and SEQ ID NO:2.

Another aspect of the invention is a method of determining the zygosityof the progeny of corn event MON89034 comprising: (a) contacting thesample comprising corn DNA with a primer set comprising SQ2842 (SEQ IDNO:6), SQ2843 (SEQ ID NO:7), SQ6523 (SEQ ID NO:10), SQ6524 (SEQ IDNO:11), PB880 (SEQ ID NO:14) and PB2931 (SEQ ID NO:15) that when used ina nucleic-acid amplification reaction with genomic DNA from corn eventMON89034, produces a first amplicon that is diagnostic for corn eventMON89034 and (b) performing a nucleic acid amplification reaction,thereby producing the first amplicon; and (c) detecting the firstamplicon; and (d) contacting the sample comprising corn DNA with saidprimer set, that when used in a nucleic-acid amplification reaction withgenomic DNA from corn plants produces a second amplicon comprising thenative corn genomic DNA homologous to the corn genomic region of atransgene insertion identified as corn event MON89034; and (e)performing a nucleic acid amplification reaction, thereby producing thesecond amplicon and (f) detecting the second amplicon; and (g) comparingthe first and second amplicons in a sample, wherein the presence of bothamplicons indicates the sample is heterozygous for the transgeneinsertion.

One aspect of the invention is providing in the diet of a lepidopteranpest an insecticidally effective amount of corn event MON89034.

Another aspect of the present invention is providing a composition orbiological sample in the form of a commodity or foodstuff that isderived from corn event MON89034, the commodity or foodstuff comprisingears of corn, shucked corn, corn silk, corn pollen, cracked corn, cornmeal, crushed corn, corn flour, corn oil, corn starch, corn steepliquor, corn malt, corn sugar, corn syrup, margarine produced from cornoil, unsaturated corn oil, saturated corn oil, corn flakes, pop corn,ethanol and/or liquor produced from corn or corn products comprising DNAdiagnostic for corn event MON89034, distillers dry goods solids (DDGS)produced from fermentation of such corn event, and animal feedscomprising such DDGS and/or corn, whether or not whole, cracked, orcrushed, processed foodstuffs, a cosmetic, and a bulking agent in whichthere is found a detectable amount of a polynucleotide that isdiagnostic for the presence of the transgenic corn event MON89034 in thebiological sample. An alternative means for providing corn as afoodstuff is to provide corn in various forms of grain for feeding, suchas whole corn, cracked corn, crushed corn, and various forms of theforegoing in a blend with milo, suet, millet, sunflower, oats, wheat,rice, beans, and the like. Detectable amounts of a nucleotide sequencein such commodity or foodstuff, such as is set forth at SEQ ID NO:1 orSEQ ID NO:2, or the complements thereof, is diagnostic for the presenceof such transgenic event MON89034 DNA in the sample, and therefore, thepresence of the transgenic event cells as having originated the DNA inthe sample.

The foregoing and other aspects of the invention will become moreapparent from the following detailed description.

DRAWINGS

FIG. 1. Organization of the transgene insert present within the genomeof transgenic corn event MON89034. The central open or white barrepresents the inserted DNA. Below the white bar is a diagram thatrepresents the various elements within the inserted DNA. The ends of theinserted DNA have been arbitrarily designated as 5′ (to the left side ofthe FIGURE) and 3′ (to the right side of the FIGURE). The Right Borderand Left Border sequences or segments are labeled beneath each end ofthe diagram illustrating the various elements within the inserted DNA.The labeled elements in the expression cassettes within the inserted DNAare, in consecutive order starting from the Right Border: e35S promoter,wheat CAB untranslated leader, rice actin intron, coding sequence forCry1A.105, wheat HSP17 3′ termination and polyadenylation sequence, FMVpromoter, hsp70 intron, rubisco small subunit chloroplast targetingpeptide coding sequence, Cry2Ab coding sequence, nos 3′ termination andpolyadenylation signal sequence, and then the Left Border. Thevertically hatched bars at either end of the central open or white barcorrespond to the arbitrarily labeled 5′ and 3′ corn genome flankingsequences. The longest black line above the hatched and open or whitebar represents SEQ ID NO:5 (the full length sequence represented by theFIGURE depicting the 5′ flanking sequence, the inserted DNA sequence,and the 3′ flanking sequence). The shorter black lines above and belowthe black line labeled as SEQ ID NO:5 represent the approximatepositions within SEQ ID NO:5 in which each of the specifically labeledsequences can be found (i.e., SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, andSEQ ID NO:4). SEQ ID NO:1 and SEQ ID NO:2, and any sequence derived fromcorn event MON89034 containing SEQ ID NO:1 and/or SEQ ID NO:2, arediagnostic for corn event MON89034 DNA in a biological sample.

DETAILED DESCRIPTION

The following definitions and methods are provided to better define thepresent invention and to guide those of ordinary skill in the art in thepractice of the present invention. Unless otherwise noted, terms are tobe understood according to conventional usage by those of ordinary skillin the relevant art. Definitions of common terms in molecular biologymay also be found in Rieger et al., Glossary of Genetics: Classical andMolecular, 5th edition, Springer-Verlag: New York, 1991; and Lewin,Genes V, Oxford University Press: New York, 1994.

As used herein, the term “corn” means Zea mays or maize and includes allplant varieties that can be bred with corn, including wild maizespecies.

As used herein, the term “comprising” means “including but not limitedto”.

A transgenic “event” is produced by transformation of plant cells withheterologous DNA, i.e., a nucleic acid construct that includes atransgene of interest, regeneration of a population of plants resultingfrom the insertion of the transgene into the genome of the plant, andselection of a particular plant characterized by insertion into aparticular genome location. The term “event” refers to the originaltransformant and progeny of the transformant that include theheterologous DNA. The term “event” also refers to progeny produced by asexual outcross between the transformant and another variety thatinclude the heterologous DNA. Even after repeated back-crossing to arecurrent parent, the inserted DNA and flanking DNA from the transformedparent is present in the progeny of the cross at the same chromosomallocation. The term “event” also refers to DNA from the originaltransformant comprising the inserted DNA and flanking genomic sequenceimmediately adjacent to the inserted DNA that would be expected to betransferred to a progeny that receives inserted DNA including thetransgene of interest as the result of a sexual cross of one parentalline that includes the inserted DNA (e.g., the original transformant andprogeny resulting from selfing) and a parental line that does notcontain the inserted DNA. The present invention relates to the eventMON89034 DNA, plant cells, tissues, seeds and processed products derivedfrom MON89034.

It is also to be understood that two different transgenic plants canalso be mated to produce offspring that contain two independentlysegregating added, exogenous genes. Selfing of appropriate progeny canproduce plants that are homozygous for both added, exogenous genes.Back-crossing to a parental plant and out-crossing with a non-transgenicplant are also contemplated, as is vegetative propagation. Descriptionsof other breeding methods that are commonly used for different traitsand crops can be found in one of several references, e.g., Fehr, inBreeding Methods for Cultivar Development, Wilcox J. ed., AmericanSociety of Agronomy, Madison Wis. (1987).

A “probe” is an isolated nucleic acid to which is attached aconventional detectable label or reporter molecule, e.g., a radioactiveisotope, ligand, chemiluminescent agent, or enzyme. Such a probe iscomplementary to a strand of a target nucleic acid, in the case of thepresent invention, to a strand of genomic DNA from corn event MON89034whether from a corn plant or from a sample that includes DNA from theevent. Probes according to the present invention include not onlydeoxyribonucleic or ribonucleic acids but also polyamides and otherprobe materials that bind specifically to a target DNA sequence and canbe used to detect the presence of that target DNA sequence.

“Primers” are isolated nucleic acids that are annealed to acomplementary target DNA strand by nucleic acid hybridization to form ahybrid between the primer and the target DNA strand, then extended alongthe target DNA strand by a polymerase, e.g., a DNA polymerase. Primerpairs of the present invention refer to their use for amplification of atarget nucleic acid sequence, e.g., by the polymerase chain reaction(PCR) or other conventional nucleic-acid amplification methods.

Probes and primers are generally 11 nucleotides or more in length,preferably 18 nucleotides or more, more preferably 24 nucleotides ormore, and most preferably 30 nucleotides or more. Such probes andprimers hybridize specifically to a target sequence under highstringency hybridization conditions. Preferably, probes and primersaccording to the present invention have complete sequence similaritywith the target sequence, although probes differing from the targetsequence and that retain the ability to hybridize to target sequencesmay be designed by conventional methods.

Methods for preparing and using probes and primers are described, forexample, in Molecular Cloning: A Laboratory Manual, 2nd ed., vol. 1-3,ed. Sambrook et al., Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y., 1989 (hereinafter, “Sambrook et al., 1989”); CurrentProtocols in Molecular Biology, ed. Ausubel et al., Greene Publishingand Wiley-Interscience, New York, 1992 (with periodic updates)(hereinafter, “Ausubel et al., 1992”); and Innis et al., PCR Protocols:A Guide to Methods and Applications, Academic Press: San Diego, 1990.PCR-primer pairs can be derived from a known sequence, for example, byusing computer programs intended for that purpose such as Primer(Version 0.5, © 1991, Whitehead Institute for Biomedical Research,Cambridge, Mass.).

Primers and probes based on the flanking DNA and insert sequencesdisclosed herein can be used to confirm (and, if necessary, to correct)the disclosed sequences by conventional methods, e.g., by re-cloning andsequencing such sequences.

The nucleic acid probes and primers of the present invention hybridizeunder stringent conditions to a target DNA sequence. Any conventionalnucleic acid hybridization or amplification method can be used toidentify the presence of DNA from a transgenic event in a sample.Nucleic acid molecules or fragments thereof are capable of specificallyhybridizing to other nucleic acid molecules under certain circumstances.As used herein, two nucleic acid molecules are said to be capable ofspecifically hybridizing to one another if the two molecules are capableof forming an anti-parallel, double-stranded nucleic acid structure. Anucleic acid molecule is said to be the “complement” of another nucleicacid molecule if they exhibit complete complementarity. As used herein,molecules are said to exhibit “complete complementarity” when everynucleotide of one of the molecules is complementary to a nucleotide ofthe other. Two molecules are said to be “minimally complementary” ifthey can hybridize to one another with sufficient stability to permitthem to remain annealed to one another under at least conventional“low-stringency” conditions. Similarly, the molecules are said to be“complementary” if they can hybridize to one another with sufficientstability to permit them to remain annealed to one another underconventional “high-stringency” conditions. Conventional stringencyconditions are described by Sambrook et al., 1989, and by Haymes et al.,In: Nucleic Acid Hybridization, A Practical Approach, IRL Press,Washington, D.C. (1985), Departures from complete complementarity aretherefore permissible, as long as such departures do not completelypreclude the capacity of the molecules to form a double-strandedstructure. In order for a nucleic acid molecule to serve as a primer orprobe it need only be sufficiently complementary in sequence to be ableto form a stable double-stranded structure under the particular solventand salt concentrations employed.

As used herein, a substantially homologous sequence is a nucleic acidsequence that will specifically hybridize to the complement of thenucleic acid sequence to which it is being compared under highstringency conditions. Appropriate stringency conditions which promoteDNA hybridization, for example, 6.0×sodium chloride/sodium citrate (SSC)at about 45° C., followed by a wash of 2.0×SSC at 50° C., are known tothose skilled in the art or can be found in Current Protocols inMolecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. Forexample, the salt concentration in the wash step can be selected from alow stringency of about 2.0×SSC at 50° C. to a high stringency of about0.2×SSC at 50° C. In addition, the temperature in the wash step can beincreased from low stringency conditions at room temperature, about 22°C., to high stringency conditions at about 65° C. Both temperature andsalt may be varied, or either the temperature or the salt concentrationmay be held constant while the other variable is changed. In a preferredembodiment, a nucleic acid of the present invention will specificallyhybridize to one or more of the nucleic acid molecules set forth in SEQID NO: 1 and 2 or complements thereof or fragments of either undermoderately stringent conditions, for example at about 2.0×SSC and about65° C. In a particularly preferred embodiment, a nucleic acid of thepresent invention will specifically hybridize to one or more of thenucleic acid molecules set forth in SEQ ID NO:1 and SEQ ID NO:2 orcomplements or fragments of either under high stringency conditions. Inone aspect of the present invention, a preferred marker nucleic acidmolecule of the present invention has the nucleic acid sequence setforth in SEQ ID NO:1 and SEQ ID NO:2 or complements thereof or fragmentsof either. In another aspect of the present invention, a preferredmarker nucleic acid molecule of the present invention shares between 80%and 100% or 90% and 100% sequence identity with the nucleic acidsequence set forth in SEQ ID NO:1 and SEQ ID NO:2 or complement thereofor fragments of either. In a further aspect of the present invention, apreferred marker nucleic acid molecule of the present invention sharesbetween 95% and 100% sequence identity with the sequence set forth inSEQ ID NO:1 and SEQ ID NO:2 or complement thereof or fragments ofeither. SEQ ID NO:1 and SEQ ID NO:2 may be used as markers in plantbreeding methods to identify the progeny of genetic crosses similar tothe methods described for simple sequence repeat DNA marker analysis, in“DNA markers: Protocols, applications, and overviews: (1997) 173-185,Cregan, et al., eds., Wiley-Liss NY; all of which is herein incorporatedby reference in its' entirely. The hybridization of the probe to thetarget DNA molecule can be detected by any number of methods known tothose skilled in the art, these can include, but are not limited to,fluorescent tags, radioactive tags, antibody based tags, andchemiluminescent tags.

Regarding the amplification of a target nucleic acid sequence (e.g., byPCR) using a particular amplification primer pair, “stringentconditions” are conditions that permit the primer pair to hybridize onlyto the target nucleic-acid sequence to which a primer having thecorresponding wild-type sequence (or its complement) would bind andpreferably to produce a unique amplification product, the amplicon, in aDNA thermal amplification reaction.

The term “specific for (a target sequence)” indicates that a probe orprimer hybridizes under stringent hybridization conditions only to thetarget sequence in a sample comprising the target sequence.

As used herein, “amplified DNA” or “amplicon” refers to the product ofnucleic-acid amplification of a target nucleic acid sequence that ispart of a nucleic acid template. For example, to determine whether thecorn plant resulting from a sexual cross contains transgenic eventgenomic DNA from the corn plant of the present invention, DNA extractedfrom a corn plant tissue sample may be subjected to nucleic acidamplification method using a primer pair that includes a primer derivedfrom flanking sequence in the genome of the plant adjacent to theinsertion site of inserted heterologous DNA, and a second primer derivedfrom the inserted heterologous DNA to produce an amplicon that isdiagnostic for the presence of the event DNA. The amplicon is of alength and has a sequence that is also diagnostic for the event. Theamplicon may range in length from the combined length of the primerpairs plus one nucleotide base pair, preferably plus about fiftynucleotide base pairs, more preferably plus about two hundred-fiftynucleotide base pairs, and even more preferably plus about fourhundred-fifty nucleotide base pairs. Alternatively, a primer pair can bederived from flanking sequence on both sides of the inserted DNA so asto produce an amplicon that includes the entire insert nucleotidesequence. A member of a primer pair derived from the plant genomicsequence may be located a distance from the inserted DNA molecule, thisdistance can range from one nucleotide base pair up to about twentythousand nucleotide base pairs. The use of the term “amplicon”specifically excludes primer dimers that may be formed in the DNAthermal amplification reaction.

Nucleic-acid amplification can be accomplished by any of the variousnucleic-acid amplification methods known in the art, including thepolymerase chain reaction (PCR). A variety of amplification methods areknown in the art and are described, inter alia, in U.S. Pat. Nos.4,683,195 and 4,683,202 and in PCR Protocols: A Guide to Methods andApplications, ed. Innis et al., Academic Press, San Diego, 1990. PCRamplification methods have been developed to amplify up to 22 kb ofgenomic DNA and up to 42 kb of bacteriophage DNA (Cheng et al., Proc.Natl. Acad. Sci. USA 91:5695-5699, 1994). These methods as well as othermethods known in the art of DNA amplification may be used in thepractice of the present invention. The sequence of the heterologous DNAinsert or flanking sequence from corn event MON89034 with seed samplesdeposited as ATCC numbers can be verified (and corrected if necessary)by amplifying such sequences from the event using primers derived fromthe sequences provided herein followed by standard DNA sequencing of thePCR amplicon or of the cloned DNA.

The amplicon produced by these methods may be detected by a plurality oftechniques. One such method is Genetic Bit Analysis (Nikiforov, et al.Nucleic Acid Res. 22:4167-4175, 1994) where an DNA oligonucleotide isdesigned which overlaps both the adjacent flanking genomic DNA sequenceand the inserted DNA sequence. The oligonucleotide is immobilized inwells of a microwell plate. Following PCR of the region of interest(using one primer in the inserted sequence and one in the adjacentflanking genomic sequence), a single-stranded PCR product can behybridized to the immobilized oligonucleotide and serve as a templatefor a single base extension reaction using a DNA polymerase and labelledddNTPs specific for the expected next base. Readout may be fluorescentor ELISA-based. A signal indicates presence of the insert/flankingsequence due to successful amplification, hybridization, and single baseextension.

Another method is the pyrosequencing technique as described by Winge(Innov. Pharma. Tech. 00:18-24, 2000). In this method an oligonucleotideis designed that overlaps the adjacent genomic DNA and insert DNAjunction. The oligonucleotide is hybridized to single-stranded PCRproduct from the region of interest (one primer in the inserted sequenceand one in the flanking genomic sequence) and incubated in the presenceof a DNA polymerase, ATP, sulfurylase, luciferase, apyrase, adenosine 5′phosphosulfate and luciferin. dNTP's are added individually and theincorporation results in a light signal which is measured. A lightsignal indicates the presence of the transgene insert/flanking sequencedue to successful amplification, hybridization, and single or multi-baseextension.

Fluorescence polarization as described by Chen, et al., (Genome Res.9:492-498, 1999) is a method that can be used to detect the amplicon ofthe present invention. Using this method an oligonucleotide is designedwhich overlaps the genomic flanking and inserted DNA junction. Theoligonucleotide is hybridized to single-stranded PCR product from theregion of interest (one primer in the inserted DNA and one in theflanking genomic DNA sequence) and incubated in the presence of a DNApolymerase and a fluorescent-labeled ddNTP. Single base extensionresults in incorporation of the ddNTP. Incorporation can be measured asa change in polarization using a fluorimeter. A change in polarizationindicates the presence of the transgene insert/flanking sequence due tosuccessful amplification, hybridization, and single base extension.

Taqman® (PE Applied Biosystems, Foster City, Calif.) is described as amethod of detecting and quantifying the presence of a DNA sequence andis fully understood in the instructions provided by the manufacturer.Briefly, a FRET oligonucleotide probe is designed which overlaps thegenomic flanking and insert DNA junction. The FRET probe and PCR primers(one primer in the insert DNA sequence and one in the flanking genomicsequence) are cycled in the presence of a thermostable polymerase anddNTPs. Hybridization of the FRET probe results in cleavage and releaseof the fluorescent moiety away from the quenching moiety on the FRETprobe. A fluorescent signal indicates the presence of theflanking/transgene insert sequence due to successful amplification andhybridization.

Molecular Beacons have been described for use in sequence detection asdescribed in Tyangi, et al. (Nature Biotech. 14:303-308, 1996) Briefly,a FRET oligonucleotide probe is designed that overlaps the flankinggenomic and insert DNA junction. The unique structure of the FRET proberesults in it containing secondary structure that keeps the fluorescentand quenching moieties in close proximity. The FRET probe and PCRprimers (one primer in the insert DNA sequence and one in the flankinggenomic sequence) are cycled in the presence of a thermostablepolymerase and dNTPs. Following successful PCR amplification,hybridization of the FRET probe to the target sequence results in theremoval of the probe secondary structure and spatial separation of thefluorescent and quenching moieties that results in the production of afluorescent signal. The fluorescent signal indicates the presence of theflanking/transgene insert sequence due to successful amplification andhybridization.

Other described methods, such as, microfluidics (US Patent pub.2006068398, U.S. Pat. No. 6,544,734) provide methods and devices toseparate and amplify DNA samples. Optical dyes used to detect andquantitate specific DNA molecules (WO/05017181). Nanotube devices(WO/06024023) that comprise an electronic sensor for the detection ofDNA molecules or nanobeads that bind specific DNA molecules and can thenbe detected.

DNA detection kits are provided using the compositions disclosed herein.The kits are useful for the identification of corn event MON89034 DNA ina sample and can be applied at least to methods for breeding corn plantscontaining the appropriate event DNA. The kits contain DNA primersand/or probes that are homologous or complementary to segments selectedfrom the sequences as set forth at SEQ ID NO:1-7, or DNA primers orprobes homologous or complementary to DNA contained in the transgenegenetic elements of DNA as set forth in the Sequence Listing. These DNAsequences can be used in DNA amplification reactions or as probes in aDNA hybridization method for detecting the presence of polynucleotidesdiagnostic for the presence of the target DNA in a sample. Theproduction of a predefined amplicon in a thermal amplification reactionis diagnostic for the presence of DNA corresponding to PTO-7455 genomeDNA in the sample. If hybridization is selected, detecting hybridizationof the probe to the biological sample is diagnostic for the presence ofthe MON89034 transgenic event DNA in the sample. Typically, the sampleis corn, or corn products or by-products of the use of corn.

The present invention provides a transgenic corn plant designated ascorn event MON89034, progeny of the plant, and cells of the plant, aswell as seed produced from the plant. Representative seed for growingthe plant, for producing progeny, for obtaining cells, or for producinga crop of said seed comprising the transgenic corn event have beendeposited on Mar. 28, 2006 with the American Type Culture Collection(ATCC) and have the accession number PTA-7455.

The plant and cells and products produced from these embodiments and thelike contain DNA that is diagnostic for the presence of DNA derived fromany cell derived from the transgenic corn event MON89034 in anybiological sample. This is because these two novel sequences arecontained within the cells of the transgenic corn event MON89034. Thediagnostic DNA comprises a nucleotide sequence that is selected from thegroup consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4,and SEQ ID NO:5. The relationship of these sequences is described moreparticularly herein and in the legend to FIG. 1 and with reference toFIG. 1.

Corn plants grown from seed that are homozygous for the DNA diagnosticfor transgenic corn event MON89034 are also within the scope of thepresent invention. Corn plants grown from seed that are heterozygous forthe DNA diagnostic for transgenic corn event MON89034 are also withinthe scope of the present invention so long as these seed also comprisethe diagnostic DNA sequences. Cells, seed, and tissue produced from suchplants comprising the diagnostic DNA are also within the scope of thepresent invention.

Corn plants, corn plant cells, pollen, ova, and the like comprising DNAdiagnostic for the transgenic corn event MON89034 exhibit resistance tolepidopteran insect infestation. These cells and plants contain DNAencoding the insecticidal protein (insecticide, toxic agent) Cry2Ab andDNA having nucleotide sequences of SEQ ID NO:1 and SEQ ID NO:2 that forma part of the genome of the cells of the plant. These plants and plantcells also contain a DNA encoding insecticidal protein (insecticide,toxic agent) Cry1A.105. These proteins can be referred to as a first anda second insecticidal protein, respectively or in the inverse.Expression of these proteins is achieved from the regulatorycomponents/genetic elements that are embedded within the expressioncassettes that provide for the expression of each of the DNA sequencesencoding these toxins and are fully described herein and in the legendto FIG. 1 and with reference to FIG. 1 and the sequence as set forth atSEQ ID NO:5. Corn plants and corn plant cells comprising these sequencesare effective for protecting plants from lepidopteran pest infestation,whether heterozygous or homozygous for the alleles in which these codingsequences are present.

The present invention also provides amplicons that can be produced fromthe sequences described herein that are diagnostic for the presence in abiological sample of DNA derived from DNA of the transgenic corn eventMON89034. An amplicon diagnostic for the presence of transgenic cornevent MON89034 DNA in a biological sample contains at least onepolynucleotide segment consisting of the nucleotide sequence as setforth at SEQ ID NO:1 or SEQ ID NO:2. These amplicons can be producedusing primer sequences as described herein below from any biologicalsample that contains at least about 0.5 femto-mole or about 0.5pico-gram of DNA derived from the transgenic corn event MON89034. Suchbiological sample sources of DNA corresponding to the transgenic cornevent MON89034 can be corn meal, corn oil, corn cake, corn seed, corngerm, corn starch, and corn flour and the like derived form thattransgenic event.

The invention also provides isolated polynucleotide molecules thatexhibit contiguous nucleotide sequences such as those set forth in SEQID NO:5. These contiguous nucleotide sequences comprise: (1) from about11 to about 12000 nucleotides and any length in-between, and furthercomprise the contiguous nucleotides as set forth at nucleotide position1-11 or 9-20 in SEQ ID NO:1 and 1-11 or 9-20 as set forth in SEQ IDNO:2; (2) any contiguous nucleotide sequence as set forth in SEQ ID NO:3from about 11 to about 2000 nucleotides and any length in-between, andfurther comprise the contiguous nucleotides as set forth at nucleotideposition 1-11 and 9-20 as set forth at SEQ ID NO:1; any contiguousnucleotide sequence as set forth in SEQ ID NO:4 from about 11 to about914 nucleotides and any length in-between, and further comprise thecontiguous nucleotides as set forth at nucleotide position 1-11 and 9-20as set forth at SEQ ID NO:2. These isolated polynucleotide molecules areuseful in DNA amplification methods to produce one or more ampliconsfrom a biological sample that contains corn DNA. The detection of suchan amplicon is diagnostic for the presence of transgenic corn eventMON89034 DNA in the sample. The isolated polynucleotide molecules arealso useful in various nucleotide detection methods for detecting thepresence of DNA derived from transgenic corn event MON89034 in abiological sample. In particular, polynucleotide probes comprising atleast about 11 contiguous nucleotide as set forth in SEQ ID NO:1 or SEQID NO:2 are useful as probes in such methods for detecting thetransgenic event MON89034 DNA in a sample. The complementary sequencesof these isolated polynucleotide molecules are also useful in the samedetection and/or amplification methods.

Kits for use in detecting the presence of DNA derived from thetransgenic corn event MON89034 in a biological sample are also provideby the present invention. A kit uses a probe polynucleotide molecule,the probe molecule containing at least from about 11 to about 12000contiguous nucleotides exhibiting substantial homology, or exhibitingsubstantial complementarity to a nucleotide segment comprising asequence as set forth at SEQ ID NO:5, would be useful for detecting thepresence of MON89034 DNA in a sample. The probe molecule should containat least one of the sequences as set forth at SEQ ID NO:1 and SEQ IDNO:2, or the complements thereof. The sequences set forth at SEQ ID NO:1and SEQ ID NO:2 are also referred to as junction sequences, i.e., thesequences within the MON89034 transgenic event genome corresponding tothe contiguous sequences at which the interrupted native genome sequenceand the transgenic DNA sequence inserted into the corn plant meet, areconnected, or are joined together. These junction sequences, arbitrarilyreferred to as 5′ and 3′ ends respectively, i.e., SEQ ID NO:1 and SEQ IDNO:2 respectively, and the complements thereof, each contain part of theinserted DNA sequence and part of the flanking corn genome sequence. Forexample, SEQ ID NO:1 represents at its 5′ half the 3′ end terminus ofthe corn genome sequence flanking the 5′ end of the inserted DNA, the 5′end of the inserted DNA being represented by the 3′ end half of thesequence as set forth at SEQ ID NO:1. SEQ ID NO:2 represents at its 5′half the 3′ end terminus of the inserted DNA, and at its 3′ end half the5′ end of the corn genome sequence flanking the 3′ end of the insertedDNA. In the naturally occurring corn genome at the position of theinserted sequence set forth in SEQ ID NO:5, the flanking sequence at the5′ end of the inserted DNA and the flanking sequence at the 3′ end ofthe inserted DNA are joined, and a first primer molecule that hybridizesto the sequence complementary to the sequence set forth at SEQ ID NO:3(other than the 3′ end 21 nucleotides of SEQ ID NO:3) and a secondprimer molecule that hybridizes to the sequence as set forth at SEQ IDNO:4 (other than the 5′ end 20 nucleotides of SEQ ID NO:4) will producean amplicon in a thermal amplification reaction with template that isDNA other than MON89034 DNA that is diagnostic for the absence of theinserted DNA in MON89034, and the same primers will produce an ampliconthat is slightly larger than 12000 nucleotides (depending on theposition of the primers in the flanking sequences set forth in SEQ IDNO:3 and SEQ ID NO:4) when using MON89034 DNA as a template. Otherembodiments are also provided.

A kit for detecting the junction sequence SEQ ID NO:1 or SEQ ID NO:2 ofcorn event MON89034 in a biological sample is provided. The kit containsa polynucleotide probe which is or is fully complementary to a sequenceselected from the group consisting of SEQ ID NO:1 or SEQ ID NO:2 orcomplements thereof, and also contains a pair of primers for use in anucleic-acid amplification reaction. The pair of primers can be referredto as a first primer consisting of at least about 15 to about 50contiguous nucleotides from the corn genome portion of SEQ ID NO:3 and asecond primer consisting of at least about 15 to about 50 contiguousnucleotides complementary to the heterologous insert DNA portion of SEQID NO:5. The first primer of the pair of polynucleotide primershybridizes specifically to the reverse complement sequence correspondingto that set forth in SEQ ID NO:3 from about nucleotide position 1through about position 2050 and the second primer of said pair ofpolynucleotide primers hybridizes specifically to the sequence as setforth in SEQ ID NO:5 from about nucleotide position 2060 through aboutnucleotide position 12,208, and are extended toward each other to forman amplicon which comprises SEQ ID NO:1, said amplicon being diagnosticfor the presence of MON89034 event DNA in the sample. A different pairof primers can be referred to as a first primer consisting of at leastabout 15 to about 50 contiguous nucleotides complementary to the corngenome portion of SEQ ID NO:4 and a second primer consisting of at leastabout 15 to about 50 contiguous nucleotides from the heterologous insertDNA portion of SEQ ID NO:5. The first primer of the pair ofpolynucleotide primers hybridizes specifically to the reverse complementsequence corresponding to that set forth in SEQ ID NO:5 from aboutnucleotide position 1 through about position 11,370 and the secondprimer of the pair of polynucleotide primers hybridizes specifically tothe sequence as set forth SEQ ID NO:4 from about nucleotide position 1through about nucleotide position 914, and are extended toward eachother to form an amplicon which comprises SEQ ID NO:2, said ampliconbeing diagnostic for the presence of MON89034 event DNA in said sample.

These primer pairs are useful in producing amplicons that compriseeither SEQ ID NO:1 or SEQ ID NO:2, as the case may be, and are thusdiagnostic for the presence of MON89034 DNA in a biological sample.These amplicons enable the detection of the presence of a junctionsequence diagnostic corn event MON89034 in a biological sample.

A method for producing and detecting an amplicon that is diagnostic fora transgenic corn event MON89034 DNA in a biological sample comprisingcorn DNA is also provided. The method comprises contacting thebiological sample together with two or more primers in a nucleic acidamplification reaction, performing a nucleic acid amplificationreaction, then detecting the amplicon. The presence of the amplicon isdiagnostic for said event DNA in the sample so long as the ampliconcontains at least one of the contiguous sequences as set forth at SEQ IDNO:1 and SEQ ID NO:2, from about nucleotide position 1-11 or 9-20, orthe complementary sequences corresponding to these positions.

The nucleotide sequences diagnostic for the presence of transgenic cornevent MON89034 in a biological sample can also be detected using othermethods. For example, contacting a biological sample suspected ofcontaining MON89034 DNA with a probe that hybridizes under stringenthybridization conditions with one or more of the nucleotide sequences asset forth at SEQ ID NO:1 or SEQ ID NO:2, subjecting the sample and probeto stringent hybridization conditions; and then detecting hybridizationof the probe to the nucleotide sequence. Detecting hybridization isdiagnostic for the presence of the MON89034 DNA in the sample.

Primer polynucleotides for use in producing, in a thermal amplificationreaction, an amplicon that is diagnostic for the presence of corn eventMON89034 DNA in a biological sample are also provided by the presentinvention. Typically, the primers are provided in pairs, the members ofthe primer pair being referred to for convenience as a first primer anda second primer. A first primer can consist of at least about 15contiguous nucleotides from the corn genome portion as set forth in SEQID NO:3 and a second primer can consist of at least about 15 contiguousnucleotides complementary to the heterologous insert DNA portion as setforth in SEQ ID NO:5. These two primers would produce an amplicon in athermal amplification reaction with template DNA obtained from cornevent MON89034 DNA that contains a polynucleotide sequence as set forthat SEQ ID NO:1. Alternatively, a first primer can consist of at leastabout 15 contiguous nucleotides from the corn genome portion of SEQ IDNO:4, and a second primer can consist of at least about 15 contiguousnucleotides complementary to the heterologous insert DNA portion of SEQID NO:5. These two primers would produce an amplicon in a thermalamplification reaction with template DNA obtained from corn eventMON89034 DNA that contains a polynucleotide sequence as set forth at SEQID NO:2.

An alternative method for detecting a junction sequence of corn eventMON89034 in a biological sample comprising corn DNA, such as SEQ ID NO:1or SEQ ID NO:2, consists of contacting the sample with a polynucleotideprobe that hybridizes under stringent hybridization conditions with oneof the junction sequences, subjecting the sample and probe to stringenthybridization conditions; and detecting hybridization of the probe tothe junction sequence. Detecting the binding/hybridization of the probeto the junction sequence is indicative of the presence of the MON89034DNA in the biological sample. A stably transformed maize plant, the DNAof which produces a DNA amplicon comprising SEQ ID NO:1 or SEQ ID NO:2when subjected to the method set forth herein, is within the scope ofthe present invention. Exemplary primer sequences, in particular, a pairof primer sequences, are set forth herein in the examples and at SEQ IDNO:6 and SEQ ID NO:7.

An alternative method of detecting the presence of corn event MON89034DNA in a biological sample can consist of the steps of contacting thesample with a probe that hybridizes under stringent hybridizationconditions with MON89034 DNA and does not hybridize under stringenthybridization conditions with corn plant genomic DNA that is notMON89034 DNA, subjecting the sample and probe to stringent hybridizationconditions, and then detecting hybridization of the probe to MON89034DNA. A probe consistent with this embodiment is or is complementary to asequence selected from the group consisting of SEQ ID NO:1 and SEQ IDNO:2. Detecting hybridization of the probe to the sample is diagnosticfor the presence of the corn event MON89034 polynucleotide in thesample. The biological sample can be any sample containing MON89034 DNAincluding but not limited to corn oil, corn meal, corn flour, corngluten, corn cakes, corn starch, corn steep liquor, corn tissue, corncells, corn grain, corn pollen, corn root tissue, DDGS, and even ethanolproduced as a byproduct of fermentation of such transgenic corn so longas the sample contains at least a detectable amount of a polynucleotidediagnostic for the presence of the MON89034 event in the sample. Apolynucleotide probe can be any nucleotide selected from the groupconsisting of a deoxyribonucleic acid, a ribonucleic acid, and anucleotide analogue, and can be labeled with at least one fluorophores,molecule containing a radio-emitting isotope, or a hapten type moleculethat can be detected specifically with an antibody or other binding typereaction.

A variety of corn comprising a DNA diagnostic for the presence of aMON89034 transgenic event DNA can be obtained by breeding a corn plantcomprising transgenic corn event MON89034 DNA together with a corn plantother than event MON89034 to produce a hybrid corn plant comprising DNAdiagnostic for said event. Such a hybrid corn plant comprising DNAdiagnostic for the transgenic corn event MON89034 is within the scope ofthe present invention, as are seed produced from the hybrid (so long asthe comprises DNA diagnostic for transgenic corn event MON89034), andpollen, ovule, seed, roots, or leaves of the hybrid corn plant MON89034,also to the extent these contain the diagnostic DNA sequences, andprogeny produced from such embodiments.

The present invention provides a method for protecting a corn plant fromlepidopteran insect infestation comprising providing in the diet of atarget lepidopteran insect pest one or more transgenic corn plant cells,each corn plant cell comprising in its genome a polynucleotidecorresponding to the sequence as set forth in both SEQ ID NO:1 and SEQID NO:2 and the contiguous nucleotide sequence as set forth in SEQ IDNO:5 between SEQ ID NO:1 and SEQ ID NO:2. The target lepidopteran insectthat feeds on such transgenic corn plant cells is inhibited from furtherfeeding on the corn plant from which the corn plant cells are derived.

Compositions are also provided by the present invention that are toxicto target lepidopteran pests of corn plants. A composition of transgenicplant cells provided in the diet of a target lepidopteran insect pest,in which each transgenic corn plant cell comprises in its genome apolynucleotide corresponding to the sequence as set forth in both SEQ IDNO:1 and SEQ ID NO:2, along with the contiguous nucleotide sequence asset forth in SEQ ID NO:5 between SEQ ID NO:1 and SEQ ID NO:2, iseffective for providing protection against lepidopteran insectinfestation to a corn plant or corn plant cell, so long as the cornplant or cell is expressing Cry1A.105 and/or Cry2Ab2 from the expressioncassettes contained within the contiguous nucleotide sequence. Suchcompositions, in the form of transgenic corn seed, have been depositedwith the American Type Culture Collection under accession numberPTA-7455. Such insect resistant corn plants, or parts thereof, willcontain DNA in the genome of the cells of such plant that have at leastone nucleotide sequence selected from the group consisting of SEQ IDNO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and SEQ ID NO:5. Progenyand seed of the insect resistant corn plant, in which the progeny andseed have the diagnostic sequences referred to herein, are also includedwithin the scope of the present invention. Such insect resistant cornplants can be produced in a method comprising crossing a transgenic cornplant event MON89034 with a different corn plant, and selecting insectresistant progeny by analyzing for at least one nucleotide sequenceselected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ IDNO:3, SEQ ID NO:4 and SEQ ID NO:5.

The insect resistant transgenic corn event MON89034 can be combined withother transgenic varieties of corn, such as corn resistant to herbicidessuch as glyphosate, glufosinate, and diacamba, and the like, or cornresistant to root devouring insects as a result of insertion ofsequences encoding proteins such as PS149B1 and modified Cry3Bb, orother varieties of transgenic corn resistant to lepidopteran insectinfestation as a result of insertion of sequences encoding other toxinproteins such as VIP3A, Cry1Ab, and Cry1Fa and the like. Variouscombinations of all of these different transgenic events are bredtogether with the corn plants of the present invention, i.e., theMON89034 event, to provide improved varieties of hybrid transgenic cornresistant to coleopteran and lepidopteran infestation, and resistant toselective herbicides. Such varieties exhibit improved yield and droughttolerance characteristics compared to non-transgenic and individualtrait transgenic varieties.

A method of producing a corn plant resistant to insect infestation isprovided, wherein the corn plant comprises an insecticidally effectiveamount of the toxin coding sequences as set forth in SEQ ID NO:5. Themethod comprises extracting the toxin coding sequences from transgeniccorn event MON89034 and introducing these coding sequences, alone ortogether, into one or more corn cells, to produce transgenic corn cellscomprising these one or more toxin coding sequences. The transgenic corncells are then grown (regenerated) into transgenic corn plantscomprising the one or more coding sequences, and the transgenic plantsthen exhibit resistance to insect infestation.

A method for determining the zygosity of the DNA of a transgenic cornplant comprising corn event MON89034 DNA, with respect to the DNA thatis diagnostic for the presence of such MON89034 DNA in a biologicalsample, is provided by the present invention. The method consists of, asa first step, contacting the sample with three different primerscomprising SEQ ID NO:6, SEQ ID NO:7, and SEQ ID NO:10, that when usedtogether in a nucleic-acid amplification reaction comprising corn eventMON89034 DNA, produces a first amplicon that is diagnostic for cornevent MON89034, and when used in a nucleic-acid amplification reactioncomprising corn genomic DNA other than MON89034 DNA, produces a secondamplicon that is diagnostic for corn genomic DNA other than MON89034DNA. The following steps consist of performing a nucleic acidamplification reaction, and comparing the amplicons produced during thethermal amplification reaction. Detecting the presence of both ampliconsis diagnostic of the zygosity of the sample. Detecting only the firstamplicon is indicative of the sample containing only MON89034 DNA, i.e.,a homozygous sample. Detecting only the second amplicon is indicative ofthe sample containing no MON89034 DNA. Detecting both the first andsecond amplicons together in a sample is indicative of a samplecontaining (1) heterozygous DNA with reference to a pure samplecontaining only heterozygous starting material, or (2) a samplecontaining both homozygous and heterozygous starting sample DNA's, or(3) a sample containing some combination of homozygous, heterozygous,and/or samples other than MON89034 DNA.

The invention also provides for growing corn plants comprising DNAdiagnostic for a transgenic DNA segment inserted into the genome of thecells of the corn plants. The DNA in the genome of the corn cellscomprises any one or all of the sequences selected from the groupconsisting of:

(a) the nucleotide sequence as set forth in SEQ ID NO:5;

(b) both of the nucleotide sequences as set forth SEQ ID NO:1 and SEQ IDNO:2;

(c) the nucleotide sequence as set forth at SEQ ID NO:3; and

(d) the nucleotide sequence as set forth at SEQ ID NO:4.

The following examples are included to demonstrate examples of certainpreferred embodiments of the invention. It should be appreciated bythose of skill in the art that the techniques disclosed in the examplesthat follow represent approaches the inventors have found function wellin the practice of the invention, and thus can be considered toconstitute examples of preferred modes for its practice. However, thoseof skill in the art should, in light of the present disclosure,appreciate that many changes can be made in the specific embodimentsthat are disclosed and still obtain a like or similar result withoutdeparting from the spirit and scope of the invention.

EXAMPLES Example 1

This Example illustrates the construction and molecular characterizationof transgenic corn event MON89034.

The corn plant MON89034 was produced by an Agrobacterium mediatedtransformation process of an inbred corn line with the plasmid constructpMON38850 (the expression cassette is shown in FIG. 1). Thetransformation method used is similar to that described in U.S. Pat. No.6,603,061. The plasmid construct pMON38850 contains the linked plantexpression cassettes with the regulatory genetic elements necessary forexpression of the Cry1A.105 insecticidal protein in corn plant cells.Corn cells were regenerated into intact corn plants consisting of atleast about 23,000 different transgenic events. Individual transgenicevents (plants) were selected from the population of events that showedintegrity of the plant expression cassettes and resistance toLepidopteran insect larvae feeding damage. A corn plant that contains inits genome the linked plant expression cassettes of pMON38850 is anaspect of the present invention. After substantial analysis of thesetransgenic events, the MON89034 transgenic event was selected on thebasis of its molecular characterization and the absence of anyundesirable phenotypic or agronomic deficiency effects.

The sequences of the transgene genetic elements contained in MON89034corn genome as illustrated in FIG. 1 consists of the following elementseach in operable linkage to each other. First at the arbitrarily defined5′ end of the sequence (i.e., near the left central portion of thesegment depicted in FIG. 1) is labeled a portion of the right borderregion (RB) from Agrobacterium tumefaciens. This is followed in sequenceby an expression cassette consisting of an enhanced CaMV 35S promoterelement (herein referred to as P-CaMV35Sen, located at positions 2350 to2651 on SEQ ID NO:5); a wheat chlorophyll A/B binding proteinuntranslated leader sequence (herein referred to as L-Ta.lhcb1, locatedat positions 2678 to 2738 on SEQ ID NO:5); a rice actin intron sequence(herein referred to as I-Os.Act1, located at positions 2755 to 3234 onSEQ ID NO:5); a non-naturally occurring sequence encoding the chimericgene Cry1A.105 (located at positions 3244 to 6777 on SEQ ID NO:5); and a3′ termination region from wheat (herein referred to asT-Ta.Hsp17-1:1:1, located at positions 6809 to 7018 on SEQ ID NO:5). Thecombination of the above referenced elements, other than the bordersequence, function together when in a corn plant to cause the expressionof the Cry1A.105 insecticidal protein. These elements are then linked insequence to another expression cassette consisting of the followingelements: a Figwort mosaic promoter (located at positions 7086 to 7649on SEQ ID NO:5), a Zea mays Hsp70 leader (herein referred to as HSP70 orI-Hsp70, located at positions 7672 to 8475 on SEQ ID NO:5), and a Zeamays chloroplast transit peptide coding sequence (herein referred to asCTP2 or TS-SSU-CTP, located at positions 8492 to 8892 on SEQ ID NO:5).These operably linked segments are then linked to a nucleotide sequenceencoding the insecticidal protein Cry2Ab (located at positions 8893 to10800 on SEQ ID NO:5), which is linked at its 3′ end to a 3′non-translated region of the nopaline synthase gene of Agrobacteriumtumefaciens (herein referred to as T-AGRtu.nos-1:1:13, located atpositions 10827 to 11377 on SEQ ID NO:5). These elements flanking theCry2Ab coding sequence function together to direct the expression ofCry2Ab when present in a corn plant. The Cry2Ab expression cassette isthen followed in sequence by a nucleotide sequence consisting of asufficient portion of the left border (LB) region from Agrobacteriumtumefaciens.

DNA molecules useful as primers in DNA amplification methods can bederived from the sequences of the genetic elements of the transgeneinsert contained in the MON89034 event. These primer molecules can beused as part of a primer set that also includes a DNA primer moleculederived from the genome of event flanking the transgene insert.

The portion of the pMON38850 plasmid DNA inserted into the corn genome,giving rise to the transgenic corn plant event MON89034, consisting ofthe left and right border segments and the two linked plant expressioncassettes (a first expression cassette encoding Cry1A.105, and a secondexpression cassette encoding Cry2Ab, wherein each cassette can beinterchangeable as to whether one is designated as being a first or asecond cassette) in between the border segments was characterized bydetailed molecular analyses. These analyses were conducted to identifyevents that contained only a single and intact inserted segmentconsisting of the borders and the desired two expression cassettes inbetween the borders (number of integration sites within the corngenome), the copy number (the number of copies of the T-DNA within onelocus), and the integrity of the inserted gene cassettes (i.e., absenceof any rearrangements or sequence variation from the sequence known tobe present in the plasmid pMON38850). DNA molecular probes were usedthat included the intact Cry1A.105 coding region and its respectiveregulatory elements, the promoters, introns, and polyadenylationsequences of the plant expression cassettes, and plasmid pMON38850backbone DNA region. The data obtained from the analyses of all eventsdemonstrated that MON89034 contains a single T-DNA insertion with onecopy of the Cry1A.105 expression cassette. No additional elements fromthe transformation vector pMON38850, linked or unlinked to intact genecassettes, were detected in the genome of MON89034. Finally, PCR and DNAsequence analyses were performed to determine the 5′ and 3′insert-to-plant genome junctions, confirm the organization of theelements within the insert (see for example, FIG. 1), and determine thecomplete sequence of the DNA inserted into the corn plant genome thatgave rise to the transgenic corn event MON89034. The complete insertedsequence, together with a portion of the corn genome flanking sequencesat either end of the inserted DNA, is depicted in the sequence as setforth at SEQ ID NO:5.

Genomic DNA from MON89034, and non transgenic DNA from corn other thanMON89034 (control DNA) was extracted from corn seed by first processingthe seed (up to 200 seeds) to a fine powder in a Harbil 5G-HD paintshaker (Harbil Inc, Cincinnati, Ohio). Briefly, the powdered seed wasextracted in extraction buffer (EM Science Cat. No. 3700, EM Science,Gibbstown, N.J., USA) and DNA precipitated from solution withisopropanol (Sigma Cat. No. 1-0398, Sigma, St. Louis, Mo., USA). Theprecipitated DNA was spooled into a microcentrifuge tube containing 70percent ethanol. The DNA was pelleted in a microcentrifuge at maximumspeed (˜14,000 rpm) for ˜5 minutes, vacuum-dried, and re-dissolved in TEbuffer (pH 8.0). The DNA was then stored in a 4° C. refrigerator. Thismethod can be modified by one skilled in the art to extract DNA from asingle corn seed.

Exemplary methods used to identify event MON89034 in a sample aredescribed in an event specific endpoint Taqman PCR for which examples ofconditions are described in Table 1 and Table 2. The DNA primers used inthe assay are primers SQ2842 (SEQ ID NO:6), SQ2843 (SEQ ID NO:7), 6FAM™labeled primer PB880 (SEQ ID NO:14) and VIC™ labeled primer PB2931 (SEQID NO:15), 6FAM and VIC are florescent dye products of AppliedBiosystems (Foster City, Calif.) attached to the DNA primers. For TaqmanMGB probes, the 5′-exonuclease activity of Taq DNA polymerase cleavesthe probe from the 5′-end, between the fluorophore and quencher. Whenhybridized to the target DNA strand, quencher and fluorophore areseparated sufficiently in three dimensional space to produce afluorescent (fluorophore excitation wavelength) signal.

SQ2842 (SEQ ID NO:6), and SQ2843 (SEQ ID NO:7), when used in thesereaction methods with PB880 (SEQ ID NO:14) produce a DNA amplicon thatis diagnostic for event MON89034 DNA. The controls for this analysisshould include a positive control from corn containing event MON89034DNA, a negative control from non-transgenic corn or from transgenic cornother than event MON89034, and a negative control that contains notemplate DNA.

SQ1564 (SEQ ID NO:17) and SQ1565 (SEQ ID NO:18) when used in thesereaction methods with PB351 (SEQ ID NO:21) produce an amplicon that isdiagnostic of Cry1A.105 in MON89034.

These assays are optimized for use with an Applied Biosystems GeneAmpPCR System 9700 or Stratagene Robocycler, MJ Engine, Perkin-Elmer 9700,or Eppendorf Mastercycler Gradient thermocycler. Other methods andapparatus known to those skilled in the art that produce amplicons thatidentify the identify event MON89034 DNA is within the skill of the art.

Any probe that binds specifically to SEQ ID NO:1 or to its perfectcomplementary sequence in a biological sample and contains at least 11contiguous nucleotides as set forth in SEQ ID NO:1, or as the case maybe, the reverse complement of the sequence in SEQ ID NO:1, so long asthe binding can be detected, is diagnostic for the presence of cornevent MON89034 DNA in that sample. Any probe that binds specifically toSEQ ID NO:2 or to its perfect complementary sequence in a biologicalsample and contains at least 11 contiguous nucleotides as set forth inSEQ ID NO:2, or as the case may be, the reverse complement of thesequence in SEQ ID NO:2, so long as the binding can be detected, isdiagnostic for the presence of corn event MON89034 DNA in that sample.

Any pair of primers that is used for or designed for use in producing anamplicon from a biological sample comprising corn DNA, and the ampliconcomprises either SEQ ID NO:1 or SEQ ID NO:2, or as the case may be,comprises both sequences, is considered to be within the scope of thepresent invention. Any such amplicon comprising either SEQ ID NO:1 orSEQ ID NO:2 or both is considered, for the purposes of the inventiondisclosed herein, to be diagnostic for the presence of the corn eventMON89034 DNA in such biological sample. The following example isprovided as reference for one skilled in the art.

TABLE 1 Corn MON89034 Event Specific Endpoint Taqman PCR Step ReagentAmount Comments 1 Nuclease-free water add to10 μl final volume — 2 2XUniversal Master Mix (Applied   5 μl 1 X final Biosystems cat. #4304437)concentration 3 Primers SQ2842 (SEQ ID NO: 6), 0.5 μl 1.0 μM final andSQ2843(SEQ ID NO: 7) concentration resuspended in nuclease-free water toa concentration of 20 μM each) 4 Primer 6FAM ™ (resuspended in 0.2 μl0.2 μM final nuclease-free water to a concentration concentration of 10μM) 5 Internal Control Primer-SQ2842 0.2 μl 0.2 μM final and internalcontrol primer SQ2843 concentration 6 Extracted DNA (template): 3.0 μlDiluted in water Samples to be analyzed (individual 4-80 ng of genomicleaves) DNA Negative control 4 ng of non-transgenic corn genomic DNANegative control no DNA template (solution in which DNA was resuspended)Positive control 4 ng of genomic DNA from known event MON89034heterozygous corn Positive control 4 ng of genomic DNA from known eventMON89034 homozygous corn 7 Gently mix, add 1-2 drops of mineral oil ontop of each reaction.

The DNA amplification can be set up and conducted using any means forthermocycling, including manual manipulations or electronicallycontrolled manipulations of temperature steps and cycles. StratageneRobocycler, MJ Engine, Perkin-Elmer 9700, or Eppendorf MastercyclerGradient thermocycler or Applied Biosystems GeneAmp PCR System 9700 orMJ Research DNA Engine PTC-225 thermal cyclers have been usedsuccessfully to conduct the following cycling parameters. When runningthe PCR in the Eppendorf Mastercycler Gradient or MJ Engine, thethermocycler was cycled in the calculated mode. When using thePerkin-Elmer 9700, the cycling conditions were conducted with the rampspeed set at maximum.

TABLE 2 Zygosity assay thermocycler conditions Cycle Settings: AppliedBiosystems GeneAmp No. PCR System 9700 1 50° C.  2 minutes 1 95° C. 10minutes 10 95° C. 15 seconds 64° C.  1 minute (−1° C./cycle) 30 95° C.15 seconds 54° C.  1 minute 1 10° C. soak

Example 2

This example illustrates the identification of a corn plant comprisingDNA diagnostic for the transgenic corn event MON89034 in its genome andthe determination of the zygosity of such corn plant.

The methods used to identify heterozygous from homozygous progenycontaining event MON89034 DNA in its genome are described in a zygosityassay for which conditions are exemplified in Table 3 and Table 4. Theexemplary DNA primers used in the zygosity assay are primers SQ2842 (SEQID NO: 6), SQ2843 (SEQ ID NO:7), SQ6523 (SEQ ID NO:10), SQ6524 (SEQ IDNO:11), 6FAM™ labeled primer PB880 (SEQ ID NO:14) and VIC™ labeledprimer PB2931 (SEQ ID NO:15). As indicated above, 6FAM and VIC areflorescent dye products of Applied Biosystems (Foster City, Calif.)attached to the DNA primer.

SQ2842 (SEQ ID NO:6), SQ2843 (SEQ ID NO:7), SQ6523 (SEQ ID NO:10),SQ6524 (SEQ ID NO:11), when used together in a thermal amplificationreaction in which a biological sample containing template DNA containsDNA that is diagnostic for the presence corn event MON89034 in thesample, produces a DNA amplicon diagnostic for corn DNA other than cornevent MON89034 DNA (independent of whether the corn DNA is derived fromnon transgenic or from some other transgenic sample). Alternatively, thereaction will produce two different DNA amplicons from a biologicalsample containing DNA derived from a corn genome that is heterozygousfor the allele corresponding to the inserted DNA present in transgeniccorn event MON89034. These two different amplicons will correspond to afirst amplicon that is derived from the wild type corn genome locus, anda second amplicon that is diagnostic for the presence of corn eventMON89034 DNA. A sample of corn DNA that gives rise only to a singleamplicon corresponding to the second amplicon described for theheterozygous genome is diagnostic for the presence of corn eventMON89034 in the sample and is diagnostic for determining that the cornDNA used as template arise from a corn seed that is homozygous for theallele corresponding to the transgenic corn event MON89034 inserted DNA.The controls for this analysis should include a positive control fromhomozygous and heterozygous corn containing event MON89034 DNA, anegative control from non-transgenic corn or any other transgenicvariety of corn, and a negative control that contains no template DNA.This assay is optimized for use with a Stratagene Robocycler, MJ Engine,Perkin-Elmer 9700, or Eppendorf Mastercycler Gradient thermocycler.Other methods and apparatus known to those skilled in the art thatproduce amplicons that identify the zygosity of the progeny of crossesmade with MON89034 plants is within the skill of the art.

TABLE 3 Zygosity assay reaction solutions Step Reagent Amount Comments 1Nuclease-free water add to 5 μl final — volume 2 2X Universal Master Mix(Applied  2.5 μl 1 X final Biosystems cat. #4304437) concentration 3Primers SEQ ID NO: 6, and SEQ ID 0.05 μl 0.25 μM final NO: 7(resuspended in nuclease-free concentration water to a concentration of20 μM) 4 PB880 SEQ ID NO: 14 Primer 0.01 μl  0.4 μM final 6FAM ™(resuspended in nuclease- concentration free water to a concentration of10 μM) 5 PB2931 SEQ ID NO: 15 Primer 0.01 μl 0.15 μM final VIC ™(resuspended in nuclease-free concentration water to a concentration of10 μM) 6 RED Taq DNA polymerase 1.0 μl (recommended to 1 unit/reaction(1 unit/μl) switch pipets prior to next step) 7 Extracted DNA(template): 2.0 μl Diluted in water Samples to be analyzed (individual4-80 ng of genomic leaves) DNA Negative control 4 ng of non-transgeniccorn genomic DNA Negative control no DNA template (solution in which DNAwas resuspended) Positive control 4 ng of genomic DNA from known eventMON89034 heterozygous corn Positive control 4 ng of genomic DNA fromknown event MON89034 homozygous corn 8 Gently mix, add 1-2 drops ofmineral oil on top of each reaction.

DNA amplification in a Stratagene Robocycler, MJ Engine, Perkin-Elmer9700, or Eppendorf Mastercycler Gradient thermocycler or AppliedBiosystems GeneAmp PCR System 9700 or MJ Research DNA Engine PTC-225thermal cycler have been used successfully to conduct the followingcycling parameters. When using the Eppendorf Mastercycler Gradient or MJEngine, the cycles were conducted in the calculated mode. When using thePerkin-Elmer 9700, the cycles were conducted with the ramp speed set atmaximum.

TABLE 4 Zygosity assay thermocycler conditions^(a) No. of Cycles inTemperature Consecutive Order and Duration 1 50° C.  2 minutes 1 95° C.10 minutes 10 95° C. 15 seconds 64° C.  1 minute (−1° C./cycle) 30 95°C. 15 seconds 54° C.  1 minute 1 10° C. soak ^(a)using AppliedBiosystems GeneAmp PCR System 9700

Seed corresponding to the transgenic event MON89034 have been depositedon Mar. 28, 2006 under the Budapest Treaty with the American TypeCulture Collection (ATCC), 10801 University Boulevard, Manassas, Va.20110. The ATCC accession number or patent deposit designation isPTA-7455. The deposit will be maintained in the depository for a periodof 30 years, or 5 years after the last request, or for the effectivelife of the patent, whichever is longer, and will be replaced asnecessary during that period.

Having illustrated and described the principles of the presentinvention, it should be apparent to persons skilled in the art that theinvention can be modified in arrangement and detail without departingfrom such principles. We claim all modifications that are within thespirit and scope of the appended claims.

All publications and published patent documents cited in thisspecification are incorporated herein by reference to the same extent asif each individual publication or patent application was specificallyand individually indicated to be incorporated by reference.

What is claimed is:
 1. A recombinant DNA molecule comprising anucleotide sequence selected from the group consisting of SEQ IDNOs:1-5, and complete complements thereof.
 2. The recombinant DNAmolecule of claim 1, wherein the DNA molecule comprises SEQ ID NO:5. 3.A corn plant, plant cell, seed, or plant part comprising the recombinantDNA molecule of claim
 1. 4. The recombinant DNA molecule of claim 1,wherein said DNA molecule comprises SEQ ID NO:1 and SEQ ID NO:2.
 5. Therecombinant DNA molecule of claim 1, wherein said DNA molecule comprisesSEQ ID NO:3 and SEQ ID NO:4.
 6. The recombinant DNA molecule of claim 1,wherein said DNA molecule comprises a sequence selected from the groupconsisting of SEQ ID NOs:1-4 and complete complements thereof, andfurther comprises a nucleotide sequence comprising a Cry1A.105 codingsequence and a nucleotide sequence comprising a Cry2Ab coding sequence.7. A corn plant, seed, cell, or plant part comprising the recombinantDNA molecule of claim
 6. 8. The corn plant, seed, cell, or plant part ofclaim 7, wherein the corn plant, seed, cell, or plant part is resistantto lepidopteran insect infestation.
 9. The corn plant, seed, cell, orplant part of claim 7, wherein the corn plant or seed is a hybrid havingat least one parent comprising corn event MON89034, a representativesample of seed comprising said event having been deposited under ATCCAccession No. PTA-7455.
 10. The corn plant, seed, cell, or plant part ofclaim 7, wherein the plant part is selected from the group consisting ofpollen, ovule, flower, grain, shoot, root, stalk, silk, tassel, ear, andleaf tissue.
 11. The corn plant, seed, cell, or plant part of claim 7,further defined as a corn plant.
 12. The corn plant, seed, cell, orplant part of claim 7, further defined as a seed.